PROJECT SUMMARY/ABSTRACT It is estimated that over 10% of all American couples suffer from infertility, a topic of great importance to NICHD and cited frequently by this Institute. Professional American women & those from all developed nations increasingly delay childbearing for several years compared to the past because of career advancement and increasing social acceptability. This problem is not solely a woman?s health issue but equally affects men as only 40% of infertility is attributable solely to women, while an estimated equal 40% is attributable to the male partner who also contributes to another 20% of combined female & male causes. Currently, recombinant (r) human (h) follicle stimulating hormone (FSH) is the single largest therapeutic component of the infertility market with sales considerably in excess of $1 billion per year, of which 40-50% of that total over $500 Million per year in the much higher priced and growing US market and another 40% in the European market. Several infertility consultants to the Trophogen PIs have confirmed that rhFSH therapy for infertility is much too expensive for most American women as the cost is not covered by insurance in many states and, even if covered, there are often prohibitive out-of-pocket costs. In addition, the results with current rhFSH treatments are far from optimal with only an average response rate of 30-35% per cycle, and much worse for women over 35, especially for greatly under-reported ?poor responders?. The latter include both to conventional first line in vivo FSH stimulation performed with intrauterine insemination (IUI) as well as to more effective but costly and invasive in vitro fertilization (IVF). Poor responders often produce fewer than 5 oocytes per superovulation cycle of variable quality and may require repeated cycles up to 6 or more to achieve a full term pregnancy, if they are successful at all, with devastating emotional and financial cost to infertile couples. Moreover, rhFSH stimulation is an important component of many rapidly growing additional markets limited only by cost; elective and medical egg banking, ethical preferred sexing of IVF babies & especially important to the PIs, future disease prevention using cutting-edge entire genome screening of IVF embryos for even more than currently screened chromosomic aberrations, all known monogenic diseases and even proclivity to complex genetic and epigenetic diseases the latter using newest metabolomics techniques. To be most effective in disease prevention, all such methods also require an improved rhFSH analog to produce the largest possible number of high quality IVF embryos of which only 2 and in the future only 1 will be transferred to the uterus. Trophogen has recently designed a 3rd generation neoglycosylated, long-acting bovine FSH analog already licensed to Zoetis (formerly Pfizer Animal Health and the world?s largest animal pharma) which has performed extensive due diligence on the safety and all aspects of the superiority of this analog compared to all currently used wild-type animal FSH products. This new bovine FSH analog has recently inspired the homologous design of the hFSH analog on an hFSH template 40 amino acids different from the bovine. Using the expedited FDA 505(b)(2) NDA regulatory track for Biosuperiors and with all these attributes (potency, maximal efficacy, long action, much lower production costs than big pharma using only inexpensive internet marketing with no need for a sales force), Trophogen is confident it will still be highly profitable, pricing its vastly superior new FSH analog in the US well below that of both currently branded FSH as well as coming biosimilars to quickly assume a dominant position within the infertility treatment market in the US & globally. In the current studies, the PIs propose in Phase 1 to: establish unequivocally the superiority of the lead TR44701 molecule based on extended in vivo testing & stimulation of ovarian weight, size & number of antral follicles; to establish a master Chinese Hamster Ovary (CHO) cell bank for future good manufacturing practices (GMP) production providing high level expression of the lead FSH analog; produce & purify additional larger of the final FSH analog candidate in roller bottles or bioreactors sufficient to submit an IND using methods of production and purification optimized in preliminary data research studies; Phase 2: transfer Trophogen?s previously developed smaller scale GLP production methods and purification methods developed in Phase 1 to the GMP facility of its about-to-be selected foreign pharma partner or alternatively to an outstanding commercial GMP facility tentatively-selected with previous methods now adapted to major up-scaling for large commercial disposable bioreactors plus addition of required new viral clearance steps; obtain larger amounts of GMP material sufficient for an FDA agreed-upon Phase 1/2 combined clinical trial as well as only one more final Phase 2B clinical pathway for Biosuperiors; perform all FDA-required efficacy, specificity, stability, metabolic, pharmacokinetic, pharmacodynamic and analytic studies; Perform FDA-IND-required toxicology studies without repro-tox studies with supplemental support from NCATS BrIDGs; preparation & submission of IND. Advanced development of this vastly superior FSH analog on a low cost, Fast-track NDA for Biosuperiors using internet marketing will make rhFSH treatment for infertility & many other growing markets, for the first time, much more effective & affordable for US & women throughout the world!